Pfeiffer A
Life Sci. 1983;33 Suppl 1:203-6. doi: 10.1016/0024-3205(83)90478-2.
Sheep brain membrane opiate receptors were solubilized using two different approaches: (A) Chaps was used to solubilize [3H]buprenorphine labeled membranes. Macromolecular labeled material had a stokes radius of approximately 90 A. Treatment with phospholipase A2, but not phospholipase C, decreased the stokes radius to 50 A indicating the presence and eventual structural role of phospholipids in the solubilized opiate receptor complex. (B) Digitonin-NaCl was used for solubilization of receptors which were then labeled with [3H]diprenorphine. This complex could be chromatographed on hydroxylapatite, DEAE-sepharose and phenylboronate gels successively permitting an approximately 100-fold purification over the solubilized starting material.
(A) 使用Chaps增溶[3H]丁丙诺啡标记的膜。大分子标记物质的斯托克斯半径约为90 Å。用磷脂酶A2而非磷脂酶C处理后,斯托克斯半径降至50 Å,表明磷脂在增溶的阿片受体复合物中的存在及其最终的结构作用。(B) 使用洋地黄皂苷 - 氯化钠增溶受体,然后用[3H]二丙诺啡进行标记。该复合物可先后在羟基磷灰石、二乙氨基乙基 - 琼脂糖和苯硼酸凝胶上进行色谱分离,相对于增溶的起始材料可实现约100倍的纯化。